Tumor-Derived Extracellular Vesicles Require β1 Integrins to Promote Anchorage-Independent Growth

DeRita, Rachel M.; Sayeed, Aejaz; Garcia, Vaughn; Krishn, Shiv Ram; Shields, Christopher D.; Sarker, Srawasti; Friedman, Andrea; McCue, Peter A.; Molugu, Sudheer K.; Rodeck, Ulrich; Dicker, Adam P.; Languino, Lucia R.

doi:10.1016/j.isci.2019.03.022

Cited by 34 publications

(31 citation statements)

References 44 publications

(91 reference statements)

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“…We apologize for not being able to describe other methods for EV isolation or commercially available kits due to space constraints. With a need for transparency and consistency in the field of EV research, investigators are encouraged to report the chosen methods used for isolation and characterization, such as expression of sEV and LEV markers, nanoparticle tracking analysis [36,37] , tunable resistive pulse sensing, flow cytometry, laser tweezers Raman spectroscopy, digital ELISA, digital PCR and transmission electron microscopy [17,38,39] .…”

Section: Resultsmentioning

confidence: 99%

Methods for extracellular vesicle isolation from cancer cells

Salem¹,

Naranjo²,

Singh³

et al. 2020

CDR

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Cells are known to release different types of vesicles such as small extracellular vesicles (sEVs) and large extracellular vesicles (LEVs). sEVs and LEVs play important roles in intercellular communication, pre-metastatic niche formation, and disease progression; both can be detected cell culture media and biological fluids. sEVs and LEVs contain a variety of protein and RNA cargo, and they are believed to impact many biological functions of the recipient cells upon their internalization or binding to cell surface proteins. It has recently been established that standard isolation techniques, such as differential ultracentrifugation, yield a mixed population of EVs. However, density gradient ultracentrifugation has been reported to allow the isolation of sEVs without cellular debris. Here, we describe the most common methods used to isolate sEVs from cell culture medium, mouse and human plasma, and a new technique for isolating sEVs from tissues as well. This article also provides detailed procedures to isolate LEVs.

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Section: Resultsmentioning

confidence: 99%

Methods for extracellular vesicle isolation from cancer cells

Salem¹,

Naranjo²,

Singh³

et al. 2020

CDR

Self Cite

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“…Integrins in EVs have garnered considerable attention for their ability to connect EVs to specific sites in the extracellular matrix [92][93][94][95]. The activity of integrins in EVs has been linked to promoting metastasis of cancer cells and to pulmonary tissue destruction [76,96,97].…”

Section: Integrins and Regulators Of Integrin Binding In Evsmentioning

confidence: 99%

“…Another consequence of the fusion of EVs is the delivery of membrane components, including integrins [121,122]. In a series of studies, data has been presented that suggests that EVs deliver integrins from a cell of origin to a target cell, and that this is an element of the machinery that allows cancer cells to precondition the local environment of a site of metastasis [93,121]. With osteoclast EVs, both alphaV beta3 and alpha2 beta1 integrins have been shown to have critical regulatory functions in osteoblasts [123][124][125].…”

Section: Integrins and Regulators Of Integrin Binding In Evsmentioning

confidence: 99%

Actin and Actin-Associated Proteins in Extracellular Vesicles Shed by Osteoclasts

Holliday

Faria

Rody

2019

IJMS

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Extracellular vesicles (EVs) are shed by all eukaryotic cells and have emerged as important intercellular regulators. EVs released by osteoclasts were recently identified as important coupling factors in bone remodeling. They are shed as osteoclasts resorb bone and stimulate osteoblasts to form bone to replace the bone resorbed. We reported the proteomic content of osteoclast EVs with data from two-dimensional, high resolution liquid chromatography/mass spectrometry. In this article, we examine in detail the actin and actin-associated proteins found in osteoclast EVs. Like EVs from other cell types, actin and various actin-associated proteins were abundant. These include components of the polymerization machinery, myosin mechanoenzymes, proteins that stabilize or depolymerize microfilaments, and actin-associated proteins that are involved in regulating integrins. The selective incorporation of actin-associated proteins into osteoclast EVs suggests that they have roles in the formation of EVs and/or the regulatory signaling functions of the EVs. Regulating integrins so that they bind extracellular matrix tightly, in order to attach EVs to the extracellular matrix at specific locations in organs and tissues, is one potential active role for actin-associated proteins in EVs.

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“…Integrins carried on SEVs have been shown to supporting tumor spread and metastasis development [30][31][32][33][34][35]. Exosomes mediate cell adhesion to matrix components [36,37], In addition, a5b1 integrin found on exosomes was shown to bind fibronectin and promote cancer cell adhesion and motility [38].…”

mentioning

confidence: 99%

Inhibition of a v b 3 integrin impairs adhesion and uptake of tumor-derived small extracellular vesicles

Altei

Pachane

Santos

et al. 2020

Preprint

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Background: Extracellular vesicles (EVs) are lipid-bound particles that are naturally released from cells and mediate cell-cell communication. Integrin adhesion receptors are enriched in small EVs (SEVs) and SEV-carried integrins have been shown to promote cancer cell migration and to mediate organ-specific metastasis; however, how integrins mediate these effects is not entirely clear and could represent a combination of EV binding to extracellular matrix and cells.Methods: To probe integrin role in EVs binding and uptake, we employed a disintegrin inhibitor (DisBa-01) of integrin binding with specificity for avb3 integrin. EVs were purified from MDA-MB-231 cells conditioned media by serial centrifugation method. Isolated EVs were characterized by different techniques and further employed in adhesion, uptake and co-culture experiments.Results: We find that SEVs secreted from MDA-MB-231 breast cancer cells carry avb3 integrin and bind directly to fibronectin-coated plates, which is inhibited by DisBa-01. SEV coating on tissue culture plates also induces adhesion of MDA-MB-231 cells, which is inhibited by DisBa-01 treatment. Analysis of EV uptake and interchange between cells reveals that the amount of CD63-positive EVs delivered from malignant MDA-MB-231 breast cells to non-malignant MCF10A breast epithelial cells is reduced by DisBa-01 treatment. Inhibition of avb3 integrin decreases CD63 expression in cancer cells suggesting an effect on SEV content.Conclusion: In summary, our findings demonstrate for the first time a key role of avb3 integrin in cell-cell communication through SEVs.

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Tumor-Derived Extracellular Vesicles Require β1 Integrins to Promote Anchorage-Independent Growth

Cited by 34 publications

References 44 publications

Methods for extracellular vesicle isolation from cancer cells

Methods for extracellular vesicle isolation from cancer cells

Actin and Actin-Associated Proteins in Extracellular Vesicles Shed by Osteoclasts

Inhibition of a v b 3 integrin impairs adhesion and uptake of tumor-derived small extracellular vesicles

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